Evolutionary analysis of the APA genes in the Phaseolus genus: wild and cultivated bean species as sources of lectin-related resistance factors?

The APA (Arcelin/Phytohemagglutinin/alpha-Amylase inhibitor) gene family is composed of various members, present in Phaseolus species and coding for lectin and lectin-related seed proteins having the double role of storage and defense proteins. Here members of the APA family have been identified by immunological, functional, and molecular analyses and representative genes were sequenced in nine wild species of Phaseolus. All taxa possessed at least one member of the true lectin gene. No arcelin type sequences have been isolated from the species examined. Among the wild species studied, only P. costaricensis contained an alpha-amylase inhibitor (alpha-AI). In addition P. augusti, P. maculatus, P. microcarpus, and P. oligospermus showed the presence of the lectin-related alpha-amylase inhibitor-like (AIL) genes and alpha-AI activity. Data from Southern blot analysis indicated the presence of only one lectin gene in P. parvulus and P. filiformis, while an extensive gene duplication of the APA locus was found in the other Phaseolus species. Phylogenetic analysis carried out on the nucleotide sequences showed the existence of two main clusters and clearly indicated that lectin-related genes originated from a paralogous duplication event preceding the development of the ancestor to the Phaseolus genus. The finding of detectable alpha-AI activity in species containing AIL genes suggests that exploiting APA genes variability in the Phaseolus genus may represent a valuable tool to find new members that may have acquired insecticidal activities.

The introduction of the stilbene synthase gene enhances the natural antiradical activity of Lycopersicon esculentum mill.

Tomato (Lycopersicon esculentum) is a vegetable rich in antioxidants, such as lycopene, lutein, and zeaxanthin. Their presence is responsible for the characteristic ability of this product to inhibit the formation of reactive oxygen species, including singlet oxygen. The grapes and wines derived from grapes also contain powerful antioxidants. The antioxidant effect is derived from the polyphenols such as resveratrol and proanthocyanidin. Resveratrol is phytoalexin that is synthesized via the activation of the gene, stilbene synthase (STS). We decided to determine if the introduction of this gene into Lycopersicon esculentum Mill could modify its antioxidant activity. Using Electronic Paramagnetic Resonance (EPR) spectroscopy, which permits the detection of antiradical activity, especially *OH (hydroxyl radical), we showed that the antioxidant activity of the products, into which the gene STS had been introduced, was almost double than that of natural products and that their activity was especially pronounced due to ripening. Moreover, resveratrol concentrations in modified tomatoes were much higher than that found in the individual fruit. In the isolated hearts subjected to ischemia/reperfusion, the rats fed with modified tomato exhibited better cardiac performance, reduced myocardial infarct size and decreased number of apoptotic cardiomyocytes, and reduced oxidative stress compared to unmodified tomato or resveratrol alone indicating superior cardioprotective abilities of modified tomatoes.

Identification and isolation of lectin nucleotide sequences and species relationships in the genus Lens Miller.

Genes for lectin, a component of legume storage proteins, were identified and characterised in two lentil cultivars ( Lens culinaris ssp. culinaris) and six wild relatives. In each taxon no differences were found among the two or three lectin clones sequenced, while differences were observed among lectin genes isolated from the different taxa. All of the clones analysed contained an insert of 828 bp and showed a high similarity with the nucleotide sequence of Pisum sativum seed lectin, PSL1. The deduced amino acid lectin sequences in all taxa were 275 amino acids long, and their multiple alignment showed that most of the variation among them occurred in regions which are not important for metal- and sugar-binding. The data from Southern blot analysis indicated the presence of only one lectin gene in all Lens taxa except L. tomentosus. Phylogenetic analyses carried out on the lectin sequences showed the existence of two main clusters and clearly indicated that L. nigricans falls outside the two groups.

[Lung cystic adenomatoid malformation: our experience]. / Malformación adenomatoidea quística, nuestra experiencia.

BACKGROUND: Cystic Adenomatoid Malformation (CAM) is a rare entity in pediatrics. To know about it will lead us to its analysis, diagnosis and adequate surgical treatment. AIM: To show our experience for the diagnosis and treatment of lung (CAM). MATERIAL AND METHODS: Over a period from 1995 to 2000 twenty-one patients with (CAM) diagnosis were treated. Diagnosis was based upon clinical evaluation, laboratory and radiological findings: these include thorax X-ray, CT scan, echo Doppler and Angiography. The elective treatment was lobectomy. RESULTS: Twenty-one patients from the Children's Hospital "Sor María Ludovica" of La Plata city were retrospectively studied over a period from 1995 to 2000. Patient's average age was 3.4 years, fourteen were feminine and seven masculine. The study consisted of clinical evaluation and imaging diagnosis: thorax-x ray; CT scan, echo Doppler and Angiography. SURGICAL TREATMENT: lobectomy. Pleural drainage was left. No deaths were registered. Pathological studies confirmed the diagnosis and type of CAM. Follow up was indicated in all patients. CONCLUSIONS: The awareness of the present pathology on the part of pediatricians makes possible considerations for careful diagnostic studies and early patient referral.

Malformación adenomatoidea quística, nuestra experiencia / Lung cystic adenomatoid malformation: our experience

Introducción. La malformación adenomatoidea quística (MAQ) es una malformación de presentación poco frecuente en pediatría; su conocimiento nos llevará a su estudio y diagnóstico posterior con tratamiento quirúrgico adecuado Objetivo. Mostrar nuestra experiencia en el diagnóstico y tratamiento de la MAQ de pulmón. Material y métodos. En el período comprendido entre el año 1995 y 2000 se trataron 21 pacientes con diagnóstico de MAQ. La metodología diagnóstica se basó en la clínica, laboratorio y diagnóstico por imágen: Rx de Tórax, TAC, Ecodoppler y Angiografía. El tratamiento consistió en lobectomía. Resultados. Se analizó en forma retrospectiva en el Hospital de Niños "Sor María Ludovica" de La Plata entre el año 1995 y 2000, 21 pacientes, edad promedio: 3,4 años, 14 de sexo femenino y 7 de sexo masculino. La metodología de estudio consistió en: Evaluación clínica. Diagnóstico por imagen: Rx. de tórax, Ecodoppler, Arteriografía, TAC. Tratamiento quirúrgico: lobectomía. Se dejó en todos drenaje pleural. No se registró ningún óbito. El estudio anátomopatológico confirmó el diagnóstico y tipo de MAQ. Se realizó seguimiento de todos los pacientes. Conclusiones. El conocimiento de la patología por parte de los pediatras, indica los estudios diagnósticos y que el paciente llegue al centro de derivación sin demoras innecesarias (AU)

Lectin-related resistance factors against bruchids evolved through a number of duplication events.

Abundant lectin-related proteins found in common beans ( Phaseolus vulgaris L.) have been shown to confer resistance against the larvae of a number of bruchid species. Genes encoding for these proteins are members of the lectin multigene family, the most representative components being arcelins, phytohemagglutinins and alpha-amylase inhibitors. Arcelins have been described in seven variants, some of which are resistance factors against the Mexican bean weevil ( Zabrotes subfasciatus), a major bean predator. In this study the isolation and sequencing of arcelin genes from wild P. vulgaris genotypes, containing Arc3 and Arc7 variants, is reported, and similarities and evolutionary relationships among the seven known arcelins are described. The evolutionary analysis shows that arcelins 3 and 4 cluster together and are the most-ancient variants. A duplication event gave rise to two additional clusters, one comprising arcelins 1, 2 and 6 and separated from the cluster of arcelins 5 and 7. A multiple number of arcelin genes were found in arcelin 3 and 4 genotypes indicating that more than one type of arcelin gene may be present in the same locus. Some of these sequences are reminiscent of ancient duplication events in arcelin evolution demonstrating that arcelins have evolved through multiple duplications. A further aim of this paper was to better understand and describe the evolution of the entire lectin multigene family. Beside arcelins, a number of other types of sequences, such as putative lectins and sequences not easily classifiable, were found in genotypes containing Arc3 and Arc4. These results, together with the evolutionary analysis, indicate that lectin loci are quite complex and confirm their origin by multiple duplication events.

Bezoares: experiencia en 10 años / Bezoares: experience in 10 years

Entre enero de 1990 y diciembre de 1999 se registraron en el Hospital de NiñosùSor María Ludovicaùde La Plata,11 pacientes con edades entre 1 y 13 años,con diagnóstico de bezoar.Se hizo un análisis retrospectivo de los casos,teniendo en cuenta:edad,sexo,motivo de consulta,manifestaciones clínicas,trastornos psicológicos,métodos diagnósticos,tratamiento y evolución.El promedio de edad fue de 5.3 años,9 eran de sexo femenino y 2 de sexo masculino.La mayoría presentó trastornos psicológicos,problemas familiares y sociales.Los antecedentes signos y síntomas fueron relevantes para el diagnóstico y confirmados con los estudios por imágenes: radiografía de abdomen en todos,ecografía abdominal en 8 y seriada gastroduodenal(SGD)en 4.Diez pacientes se trataron quirurgicamente,4 ingresaron con diagnóstico de abdomen agudo oclusivo,operándose de urgencia.Uno fue un tricobezoar de localización ileal,otro un tricobezoar de localización gástrica,y 2 fitobezoares(semillas de girasol)uno tenía impactación en estenosis del ángulo esplénico del colon(secuela de símdrome urémico hemolítico)y el otro con localización rectal,tratado con extracción manual.El resto se operó en forma programada,6 de ellos fueorn de localización gástrica,y un síndrome de Rapunzel.Ocho casos fueron tricobezoares,2 fitobezoares y uno mixto.La evolucíón postoperatoria fue favorable y el seguimiento fue favorable y el seguimiento se realizó conjuntamente con el servicio de psicopatolología

Bezoares: experiencia en 10 años / Bezoares: experience in 10 years

Entre enero de 1990 y diciembre de 1999 se registraron en el Hospital de Niños¨Sor María Ludovica¨de La Plata,11 pacientes con edades entre 1 y 13 años,con diagnóstico de bezoar.Se hizo un análisis retrospectivo de los casos,teniendo en cuenta:edad,sexo,motivo de consulta,manifestaciones clínicas,trastornos psicológicos,métodos diagnósticos,tratamiento y evolución.El promedio de edad fue de 5.3 años,9 eran de sexo femenino y 2 de sexo masculino.La mayoría presentó trastornos psicológicos,problemas familiares y sociales.Los antecedentes signos y síntomas fueron relevantes para el diagnóstico y confirmados con los estudios por imágenes: radiografía de abdomen en todos,ecografía abdominal en 8 y seriada gastroduodenal(SGD)en 4.Diez pacientes se trataron quirurgicamente,4 ingresaron con diagnóstico de abdomen agudo oclusivo,operándose de urgencia.Uno fue un tricobezoar de localización ileal,otro un tricobezoar de localización gástrica,y 2 fitobezoares(semillas de girasol)uno tenía impactación en estenosis del ángulo esplénico del colon(secuela de símdrome urémico hemolítico)y el otro con localización rectal,tratado con extracción manual.El resto se operó en forma programada,6 de ellos fueorn de localización gástrica,y un síndrome de Rapunzel.Ocho casos fueron tricobezoares,2 fitobezoares y uno mixto.La evolucíón postoperatoria fue favorable y el seguimiento fue favorable y el seguimiento se realizó conjuntamente con el servicio de psicopatolología

Lectin and lectin-related proteins in lima bean (Phaseolus lunatus L.) seeds: biochemical and evolutionary studies.

Lectin-related polypeptides are a class of defence proteins found in seeds of Phaseolus species. In Lima bean (P. lunatus), these proteins and their genes have been well characterized in the Andean morphotype, which represents one of the two gene pools of this species. To study the molecular evolution of the lectin family in Lima bean we characterized the polypeptides belonging to this multigene family and cloned the genes belonging to the Mesoamerican gene pool. The latter gene pool contains components similar to those of the Andean pool, namely: an amylase inhibitor-like (AIL), an arcelin-like (ARL) lectin and the less abundant Lima bean lectin (LBL). These proteins originate from an ancestor gene of the lectin type which duplicated to yield the lectin gene and the progenitor of ARL and AIL. In this species. ARL represents an evolutionary intermediate form that precedes AIL. Phylogenetic analysis supports an Andean origin for Lima bean. The molecular evolutionary studies were extended to the genes of common bean and demonstrated that true lectin genes and the ancestor of lectin-related genes are the result of a duplication event that occurred before speciation. Lima and common bean followed different evolutionary pathways and in the latter species a second duplication event occurred that gave rise, in Mesoamerican wild genotypes, to arcelin genes.

Effects of left ventricular pacing on cardiac performance and on quality of life in patients with drug refractory heart failure.

We assessed the effects of left ventricular pacing on echocardiographic and clinical parameters in 13 consecutive patients with heart failure and bundle branch block by means of a controlled acute and medium-term evaluation. Left ventricular pacing induced a significant improvement in left ventricular ejection fraction, Minnesota Living with Heart Failure Questionnaire score, New York Heart Association class, and 6-minute walking test compared with sinus rhythm or right ventricular pacing.

Misfolding and aggregation of vacuolar glycoproteins in plant cells.

Phaseolin and lectin-related polypeptides, the abundant oligomeric glycoproteins of bean seeds, are synthesized on the endoplasmic reticulum (ER) and then transported to the storage vacuole via the Golgi apparatus. Glycosylation and folding are among the major modifications these proteins undergo in the ER. Although a recurrent role of N-glycosylation is on protein folding, in previous studies on common bean (Phaseolus vulgaris) seeds we demonstrated that the oligosaccharide side-chains are not required for folding, intracellular transport and activity of storage glycoproteins. We show here that in lima bean (Phaseolus lunatus), incubation of the developing cotyledon with tunicamycin to prevent glycosylation has a dramatic effect on the intracellular transport of the storage glycoproteins. When lacking their glycans, phaseolin and lectin-related polypeptides misfold and are retained in the ER as mixed aggregates to which the chaperone BiP irreversibly associates. The lumen of the ER becomes enlarged to accommodate the aggregated polypeptides. Intracellular transport of legumin, a naturally unglycosylated storage protein, is mostly unaffected by the inhibitor, indicating that the observed phenomenon specifically occurs on glycoproteins. Furthermore, recombinant lima bean phaseolin synthesized in tobacco protoplasts is also correctly folded and matured in the presence of tunicamycin. To our knowledge, this is the first report that describes in detail the block of intracellular transport of vacuolar glycoproteins in plant cells due to aggregation following glycosylation inhibition.

Malformacion pulmonar: secuestro / Lung malformation: sequestration

Presentamos un caso de secuestro pulmonar hallado en un lactante de 2 meses. Los estudios de imagenes (TAC y Eco Doppler)resultaron criticos para definir el diagnostico al demostrar la presencia de un vaso arterial anomalo originado en la aorta abdominal. La cirugia fue curativa

Invaginacion intestinal en la purpura de Schonlein-Henoch / Intestinal invaginacion in Schonlein-Henoch Purpura

Se presenta un caso de invaginacion intestinal en un niño de 3 años de edad con diagnostico de purpura de Schonlein-Henoch. La asociaciones particularmente importante por: a) la baja incidencia de la combinacion de estas dos entidades (1,3 al 13,6 de los casos de purpura, con solo 101 caso comunicados a la fecha),y b)la alta morbimortalidad por retraso en el diagnostico. Este resulto de la sospecha clinica oportuna yla aplicacion de tecnicas de diagnosticos y tratamiento que permitieron resolver adecuadamente el caso. La utilizacion temprana de la ultrasonografia y el procedimiento de estudio radiologicode con por enema puede evitar la intervencion quirurgica de estos pacientes

Malformacion pulmonar: secuestro / Lung malformation: sequestration

Presentamos un caso de secuestro pulmonar hallado en un lactante de 2 meses. Los estudios de imagenes (TAC y Eco Doppler)resultaron criticos para definir el diagnostico al demostrar la presencia de un vaso arterial anomalo originado en la aorta abdominal. La cirugia fue curativa

Invaginacion intestinal en la purpura de Schonlein-Henoch / Intestinal invaginacion in Schonlein-Henoch Purpura

Se presenta un caso de invaginacion intestinal en un niño de 3 años de edad con diagnostico de purpura de Schonlein-Henoch. La asociaciones particularmente importante por: a) la baja incidencia de la combinacion de estas dos entidades (1,3 al 13,6 de los casos de purpura, con solo 101 caso comunicados a la fecha),y b)la alta morbimortalidad por retraso en el diagnostico. Este resulto de la sospecha clinica oportuna yla aplicacion de tecnicas de diagnosticos y tratamiento que permitieron resolver adecuadamente el caso. La utilizacion temprana de la ultrasonografia y el procedimiento de estudio radiologicode con por enema puede evitar la intervencion quirurgica de estos pacientes

Quiste de mesenterio: forma inusual de presentación / Mesenteric cyst: unusual presentation

Los quistes mesentéricoas y retroperitoneales son formaciones derivadas del tejido ectópico linfático.Muchos en su ubicación abdominal pueden ser sintomáticos o accidentalemente encontrados en un acto quirúrgico.ero la presentación como una hernia inguinal atascada es extremadamente rara,sólo se ha descripto 5 casos en la literatura de habla inglesa

Quiste de mesenterio: forma inusual de presentación / Mesenteric cyst: unusual presentation

Los quistes mesentéricoas y retroperitoneales son formaciones derivadas del tejido ectópico linfático.Muchos en su ubicación abdominal pueden ser sintomáticos o accidentalemente encontrados en un acto quirúrgico.ero la presentación como una hernia inguinal atascada es extremadamente rara,sólo se ha descripto 5 casos en la literatura de habla inglesa

Novel lectin-related proteins are major components in lima bean (Phaseolus lunatus L.) seeds.

The only component of the lectin-related protein family so far reported in Lima bean (Phaseolus lunatus L.) seeds is the minor seed lectin (LBL). In the morphotype Big Lima, we have isolated and characterised two abundant lectin-related seed proteins and the corresponding cDNA clones. The clones show 93.7% nucleotide identity and encode an arcelin-like (ARL) and an alpha-amylase inhibitor-like (AIL) protein. Not considering the signal peptides, ARL and AIL polypeptides contain 239 and 233 amino acids, respectively. Each polypeptide is present in the mature protein as two glycoforms. ARL subunits (43 and 46 kDa) make up oligomers of about 125 to 130 kDa whereas AIL subunits (40 and 42 kDa) oligomerise in dimers of about 88 to 100 kDa. cDNA clones encoding two isoforms of the less abundant Lima bean lectin were also isolated. In common bean (P. vulgaris) the lectin locus encodes the lectin and the lectin-related proteins alpha-amylase inhibitor and arcelin, all plant defence proteins. Our data indicate extensive evolution of the locus also in Lima bean.

Protein quality control along the route to the plant vacuole.

To acquire information on the relationships between structural maturation of proteins in the endoplasmic reticulum (ER) and their transport along the secretory pathway, we have analyzed the destiny of an assembly-defective form of the trimeric vacuolar storage glycoprotein phaseolin. In leaves of transgenic tobacco, where assembly-competent phaseolin is correctly targeted to the vacuole, defective phaseolin remains located in the ER or a closely related compartment where it represents a major ligand of the chaperone BiP. Defective phaseolin maintained susceptibility to endoglycosidase H and was slowly degraded by a process that is not inhibited by heat shock or brefeldin A, indicating that degradation does not involve transport along the secretory pathway. These results provide evidence for the presence of a quality control mechanism in the ER of plant cells that avoids intracellular trafficking of severely defective proteins and eventually leads to their degradation.